非亲缘非清髓造血干细胞移植成功治疗急性淋巴细胞白血病首例报告

为探讨非亲缘非清髓异基因造血干细胞移植（URD－NAPBSCT）在治疗急性白血病中（AL）的作用，1例HLA完全相合的急性淋巴细胞白血病病人接受了URD－NAPBSCT治疗。结果病人顺利度过造血抑制期并获得完全性植入，移植后发生了皮肤Ⅱ度GVHD和间质性肺炎，经治疗后痊愈。结果初步表明，URD－NAPBSCT较简便安全，可为急性白血病提供新的有效治疗手段。     

非清髓异基因造血干细胞移植治疗重症再生障碍性贫血4例

为探讨非清髓性异基因外周血造血干细胞移植（NAPBSCT）在治疗重症再生障碍性贫血（SAA）中的作用。4例HLA完全相合的重症再生障碍贫血病人（2例肝炎相关性重症再生障碍性贫血,2例重症再生障碍性贫血）接受了NAPBSCT,结果4例均顺利度过造血抑制期并移植成功（完全性植入3例,嵌和性植入1例）,外周血象及骨髓造血功能快速恢复正常。4例均无急性移植物抗宿主病和间质性肺炎等并发症。本结果初步表明：NAPBSCT简单、安全、并发症轻、造血功能重建快,为SAA的治疗开辟了一条新途径。     

荧光原位杂交技术在膀胱癌早期诊断中的作用

目的通过荧光原位杂交(FISH)技术检测尿液内脱落细胞染色体异常情况,以提高膀胱癌早期诊断率,减少患者膀胱镜检查次数。方法应用FISH技术分析110例血尿患者尿脱落细胞染色体异常情况,其中包括膀胱癌86例、泌尿系感染12例、滤泡增生(腺性膀胱炎)4例、良性肿瘤(乳头状瘤)4例、不明原因血尿4例。上述患者均经膀胱镜组织活检确诊。比较FISH检测与病理切片检查在确诊膀胱肿瘤上的区别。结果 86例膀胱癌患者中有68例FISH检测结果为阳性,剩余18例FISH检测为阴性的膀胱癌患者中有10例为小于5mm的Ta期浅表膀胱癌。与病理学检查结果比较,FISH检测的特异性为100%,敏感性为87%。FISH检测结果发现T1及T2期膀胱癌细胞染色体异常数目明显高于Ta期膀胱肿瘤(P<0.05),但在T1及T2期膀胱肿瘤间差异无统计学意义(P>0.05)。结论 FISH检测具有敏感性高、特异性强、无创伤等优点,在初步诊断及监测膀胱癌复发方面具有较好的临床应用前景,有可能成为简便、有效的膀胱癌初筛指标之一。     

应用早熟染色体凝聚方法研究电离辐射对细胞的损伤及修复

大量的实验已证明电离辐射诱发的淋巴细胞染色体畸变可作为可靠的生物剂量仪。然而常规细胞遗传学方法,只能分析分裂中期细胞中的染色体,间期的淋巴细胞要进入有丝分裂期,必须在植物血凝素刺激下培养二天才能达到。     

供者同后造血干细胞输注在非清髓异基因造血干细胞移植中的应用

为探讨供者造血干细胞输注 (DSI)在非清髓异基因外周血干细胞移植 (NAPBSCT)中的作用 ,6例NAPBSCT患者分别在移植后 +7～ +90天进行DSI治疗 ,共输注单个核细胞 (MNC) (0 .6～ 7.6 )× 10 8/kg ,CD34 + 细胞 (0 .3～ 3.4)× 10 6/kg ,CD3+ 细胞 (0 .3～ 5 .1)× 10 8/kg。 6例中 5例有明显促进植入的作用 ,其中 3例由嵌合体转为完全性植入。 4例有较明确的移植物抗白血病效应 (GVL) ,均未见造血抑制等并发症。初步结果表明 ,DSI能促进NAPBSCT后供者嵌合体的增加 ,有利于NAPBSCT后供者细胞植入并具较好的GVL效应 ,而且并发症少 ,在NAPBSCT中具有较好的应用前景。     

荧光原位杂交技术在流产胚胎染色体检查与产前诊断中应用价值研究

目的探讨荧光原位杂交技术(FISH)在流产胚胎染色体检查与产前诊断中的临床应用价值。方法选取自2015年1月至2017年1月南通市妇幼保健院收治的自然流产孕妇279例,产前诊断并行羊膜穿刺检查的孕妇57例。采集流产胚胎绒毛组织、产前诊断羊水,分别通过常规方法(细胞培养+G显带分析)与FISH分析核型。结果 279例流产胚胎绒毛组织标本中,常规方法检测完成87例(31.18%),检出染色体异常11例(12.64%);FISH检测完成279例(100.00%),检出染色体异常104例(37.28%),其中,常染色体数目异常占59.62%(62/104),多倍体异常占18.27%(19/104),性染色体数目异常占5.77%(6/104),复合异常占4.81%(5/104),其他异常占11.53%(12/104)。两种方法可检概率与异常结果检出概率比较,差异均有统计学意义(P<0.05)。两种方法检测结果一致概率为88.51%(77/87)。57例产前诊断羊水标本中,常规方法检测完成57例(100.00%),检出染色体异常3例(5.26%);FISH检测完成57例(100.00%),检出染色体异常3例(5.26%)。两种方法可检概率与异常结果检出概率比较,差异均无统计学意义(P>0.05)。两种方法检测结果一致概率为100.00%(57/57)。结论 FISH在流产胚胎染色体检查与产前诊断中优势明显,可作为常规方法的补充与替代。     

用DSCR Cosmid DNA探针快速基因诊断先天愚型

目的　采用DSCRCosmidDNA探针作为检测先天愚型的特异性探针。方法　对 46例先天愚型患者外周血淋巴细胞分别在经过细胞培养和不经细胞培养两种处理条件下同时进行FISH检测 ,结果　在中期染色体和间期细胞核中观察到 3个杂交信号 ,杂交率均为 81%～ 95 %。结论　表明DSCRCosmidDNA探针可直接在未经培养的淋巴细胞间期核中进行FISH检测。 48h内能对先天愚型患者做出快速、准确的诊断。     

巴西格埃尼亚事故7.5年后~(137)Cs受照者的淋巴细胞染色体易位

目的:检测137　Cs事故受照7.5年后淋巴细胞染色体易位,并与事故后初始双着丝粒频率相比较,探讨用于辐射剂量重建的基础。方法:12例格埃尼亚事故受照者,在1995年3月(事故后7.5年)取血进行细胞遗传学检查,事故后估算的受照剂量为0.2～4.6Gy。外周血样在RPMI1640培养液中培养48h,用1、6、11和3、4、8号染色体两组探针进行荧光原位杂交(FISH)方法分析,同时进行常规方法全染色体畸变分析。取12例未受照者的淋巴细胞培养作对照。结果:对12例事故受照者用吉姆萨染色中期细胞,染色体总畸变率100个细胞变动在0.8～6.8之间,双着丝粒 环为0～2.8;与…     

高海拔地区FISH法检测乳腺癌HER2基因的方法改进

目的 在高海拔地区改良并简化乳腺癌HER2 FISH法检测的预处理步骤,节省实验时间,扩大FISH检测试剂盒的临床应用.方法 以10例HER2免疫组化染色结果为＋ ＋以上的乳腺癌标本为研究对象,采用美国PanPath公司的HER2 FISH检测试剂盒,将其中的预处理步骤改良为高压煮沸处理,比较常规法与改良法对HER2基因扩增结果的影响.结果 两种方法检测乳腺癌HER2基因的阳性率无统计学差异,并且均能使细胞间质完全消化,细胞或重叠或孤立,细胞核中红绿信号清晰.结论 改良FISH法可以替代常规预处理步骤,简化了步骤,节省了时间,扩大了FISH检测试剂盒的临床应用.     

052 用FISH测定原子弹爆炸幸存者的稳定染色体畸变：与以前用Giemsa染色的相同230例资料比较

目的：评价常规Giemsa染色和FISH（荧光原位杂交）方法测定原子弹爆炸幸存者外周血淋巴细胞染色体畸变的效能。方法：血样采自２３０例广岛原子弹爆炸幸存者，４０～８０岁，按１９８６剂量系统估算的平均骨髓剂量为０．８２Sv。１９７０～１９９３年间取血用常规Giemsa染色方法和１９９２～１９９６年间取血用FISH方法测定染色体畸变，前者每个血样检测１００个细胞，染色体畸变分为３型：相互易位、臂间倒位和微小缺失；FISH分析用异硫氰基荧光素和碘化丙啶标记的１、２和４号染色体复合探针原位杂交，每个血样检测５００～６００个…     

非清髓异基因造血干细胞移植治疗急性白血病的临床研究

采用非清髓预处理（环胞霉素A、环磷酰胺、阿糖胞苷及CD3单克隆抗体或抗淋巴细胞球蛋白）的异基因外周造血干细胞移植治疗急性白血病5例。5例病人均顺利度过造血抑制期并移植成功。2例例经嵌合性植入转为完全植入,另3例为嵌合性植入。5例中2例发生Ⅱ～Ⅲ度GVHD,经治疗后治愈。5例中4例仍无病存活。结果表明,非清髓异基因造血干细胞移植简便安全,并发症少,疗效较好,为急性白血病的治疗提供了新手段。     

异基因外周血造血干细胞移植后造血细胞嵌合状态的分析

为评估异基因外周血造血干细胞移植（ALLO－PBSCT）后的植入状况，采用细胞遗传学分析结合短串联重复序列－聚合酶链反应（STR－PCR），对30例Allo-PBSCT病人进行了1－25个月的随访观察。结果表明，不同性别移植的23例，21例完全嵌合（CC），1例混合嵌合（MC），1例为完全受者核型；相同性别移植的慢性髓性白血病（CML）7例，除2例外，其余均未检测到Ph染色体；STR－PCR检测结果与染色体分析结果完全一致。说明两种方面具有互补性，可结合使用。     

亲缘间HLA半相合外周血造血干细胞移植治疗急性白血病的临床研究

目的探讨HLA半相合外周血造血干细胞移植(PBSCT)治疗急性白血病的疗效及安全性。方法4例急性白血病患者行HLA不全相合PBSCT,其中急性淋巴细胞白血病2例,急性非淋巴细胞白血病-M4b1例、M4EO1例,移植时均处于完全缓解状态。HLA配型1、2个位点不合各1例,3个位点不合2例。预处理方案由全身照射或白消胺、阿糖胞苷、环磷酰胺、司莫司汀组成。移植物抗宿主病(GVHD)的预防采用环胞素A、甲氨蝶呤、霉酚酸酯、抗胸腺细胞球蛋白四联方案。结果患者移植过程顺利,4例患者均获得造血功能重建并达完全供者植入。3例出现Ⅰ度急性GVHD,1例出现Ⅱ度急性GVHD。1例并发巨细胞病毒间质性肺炎,2例并发真菌肺炎。至今2例患者分别无病存活30月和15月,1例目前为移植后4月,1例死亡。结论HLA半相合PBSCT疗效较好、安全可行,为无HLA完全相合供者的白血病患者提供了新的治疗手段。     

军事作业人员对图片性信息敏感度的内隐实验研究

通过图片内隐实验的方法对 4个兵种的 44 8名军事作业人员和 86名地方被试进行了性信息敏感度的研究。通过预实验筛选出 2 0对性信息敏感度测试图片 ,然后以每对 5 0 0ms的速度快速呈现配对图片 ,研究被试对性信息图片和无性信息图片选择的差异。结果发现 ,与地方被试相比 ,4组军事作业人员都表现出明显的选择性信息图片的倾向性 ,但这种倾向性未发现存在性别和组间差异。本研究提示 ,与地方被试比较 ,军事人员对于性信息具有更高的敏感度 ;图片内隐实验可为社会认知的性信息敏感度研究提供一种新的手段     

Detection of stable chromosome aberrations by FISH in A-bomb survivors: comparison with previous solid Giemsa staining data on the same 230 individuals

PURPOSE: To evaluate the relative abilities of the solid Giemsa staining (conventional) and fluorescence in situ hybridization (FISH) methods in the detection of stable chromosome aberrations in the peripheral blood lymphocytes of A-bomb survivors. MATERIALS AND METHODS: Lymphocytes from a total of 230 A-bomb survivors for whom prior chromosome aberration data had been obtained by the conventional method were recently examined afresh using FISH in which chromosomes 1, 2 and 4 were painted with composite probes. RESULTS: It was found that the early use of the solid Giemsa staining method had allowed the detection of translocations with a mean frequency of 73% of the value for the genome-equivalent translocation frequency (F(G)) that was now obtained using FISH. The disparity may at least in part be due to the reciprocal exchange of seemingly identical amount of chromosome material; such exchanges can escape detection by the conventional method but can be readily identified using FISH. CONCLUSION: It has previously been established that the conventional method can detect about 20% of radiation-induced translocations as abnormal monocentric chromosomes. Present results indicate that an additional 50% can be detected if proper karyotyping is conducted and the remaining 30% are not likely to be detected unless FISH or banding methods are used. Thus, solid Giemsa staining accompanied by karyotyping may not be quite as unsuitable as is generally assumed for retrospective biodosimetry analyses, which deal mainly with stable aberrations.     

Detection of stable chromosome aberrations by FISH in A-bomb survivors: comparison with previous solid Giemsa staining data on the same 230 individuals

Purpose : To evaluate the relative abilities of the solid Giemsa staining (conventional) and fluorescence in situ hybridization (FISH) methods in the detection of stable chromosome aberrations in the peripheral blood lymphocytes of A-bomb survivors. Materials and methods : Lymphocytes from a total of 230 A-bomb survivors for whom prior chromosome aberration data had been obtained by the conventional method were recently examined afresh using FISH in which chromosomes 1, 2 and 4 were painted with composite probes. Results : It was found that the early use of the solid Giemsa staining method had allowed the detection of translocations with a mean frequency of 73% of the value for the genome-equivalent translocation frequency (F G) that was now obtained using FISH. The disparity may at least in part be due to the reciprocal exchange of seemingly identical amount of chromosome material; such exchanges can escape detection by the conventional method but can be readily identified using FISH. Conclusion : It has previously been established that the conventional method can detect about 20% of radiation-induced translocations as abnormal monocentric chromosomes. Present results indicate that an additional 50% can be detected if proper karyotyping is conducted and the remaining 30% are not likely to be detected unless FISH or banding methods are used. Thus, solid Giemsa staining accompanied by karyotyping may not be quite as unsuitable as is generally assumed for retrospective biodosimetry analyses, which deal mainly with stable aberrations.     

Sex determination using discriminant function analysis in children and adolescents: a lateral cephalometric study

The objective of this study is to test the validity of sex determination in children and adolescents using lateral radiographic cephalometry and discriminant function analysis. Fifty male and 50 female cephalograms of Taiwanese children were used (males and females with mean age of 15.52 ± 1.38 and 15.67 ± 1.54 years, respectively). Twenty-two cephalometric measurements were performed using computerized cephalometry. Statistical analysis shows that all measurements were sexually dimorphic (p < 0.05). Nine measurements, statistically validated and clinically relevant, were used for discriminant function analysis. A stepwise discriminant procedure selected seven of the nine variables, producing 95% accuracy in sex determination. Resubstitution classification reveals the same discriminant rate. Cross-validation classification (the leave-one-out method) reveals that the correct sex determination rate is 91%. However, the combination of four variables using both the stepwise procedure and the resubstitution method achieves a 92% accuracy rate. A cross-validation classification procedure with the same four variables resulted in a 91% accuracy rate. Therefore, this study uses four cephalometric measurements as the minimum number of traits yielding the maximum discriminant effectiveness of sex determination in children and adolescents.     

荧光染色体原位杂交技术在产前诊断中的应用

目的 建立并优化荧光染色体原位杂交技术并评价其在胎儿21、18、13、X和Y染色体数目异常产前诊断中的临床应用价值.方法 常规穿刺取羊水或脐带血样本196例,经低渗、固定、制片、老化等,直接利用两组特异性探针(GLP 13/GLP 21和CSP18/CSP X/CSP Y)进行原位杂交,快速检测羊水或脐带血细胞中21、18、13、X和Y染色体的数目,并同时对样本进行经典的细胞培养和染色体核型分析.结果 以≥90％的细胞显示的荧光信号点数目作为染色体数目的判断标准,正常情况下GLP 13/GLP 21探针组显示2个绿色荧光点/2个红色荧光点,CSP18/CSP X/CSPY探针组显示2个蓝绿色荧光点/2个黄色荧光点(女性)或者2个蓝色荧光点/1个黄色荧光点/1个红色荧光点(男性).196例羊水或脐血均在72～96h内给出报告,检出21三体综合征7例、l8三体综合征2例、性染色体异常(47,XYY)1例,与1个月后报告的染色体核型分析结果一致.结论 荧光染色体原位杂交方法可用于快速产前诊断胎儿21、18、13、X、Y染色体数目异常,与常规的染色体核型分析技术结合,可快速准确地检测胎儿染色体的非整倍体异常.     

Evaluation of spectral karyotyping (SKY) in biodosimetry for the triage situation following gamma irradiation

PURPOSE: Biological dosimetry in an acute triage situation of radiation exposure is traditionally performed by scoring unstable dicentric chromosomal aberrations after conventional Giemsa staining, and more recently also by detection of chromosomal translocations after chromosome painting by fluorescence in situ hybridization (FISH). By spectral karyotyping (SKY) each chromosome can be painted in an individual colour, permitting the scanning for structural aberrations throughout the genome in each individual metaphase. Here we have evaluated the performance of SKY analysis in a simulated triage situation after gamma irradiation. MATERIALS AND METHODS: Peripheral leukocytes were irradiated by 60Co (0 - 5 Gy) and analysed by SKY, Giemsa staining and FISH painting of chromosomes 1, 2, and 3. RESULTS: At 1 Gy and higher doses, dicentric aberrations (Dic+) as well as classical one- and two-way translocations were found in increasing and dose-dependent frequencies by SKY. The frequency of dicentrics detected by Giemsa was found to be significantly higher than the total aberrations detected by SKY (p<0.001), but did not differ significantly from that of FISH painting. The difference was mainly attributable to the low sensitivity of SKY to detect Dic+ following frequent lack of acentric fragments with matching chromosomal composition. CONCLUSIONS: The findings anticipate that radiation induced chromosomal aberrations may be more complex than expected from conventional and single chromosome painting analyses. While conventional Giemsa staining was found to be the method of choice for the triage situation, it is expected that extended SKY analysis will add to the knowledge of underlying mechanisms for irradiation associated chromosomal aberrations.     

Rapid translocation frequency analysis in humans decades after exposure to ionizing radiation.

This paper presents an analysis of the utility of fluorescence in situ hybridization (FISH) with whole-chromosome probes for measurement of the genomic frequency of translocations found in the peripheral blood of individuals exposed to ionizing radiation. First, we derive the equation: Fp = 2.05fp(1-fp)FG, relating the translocation frequency, Fp, measured using FISH to the genomic translocation frequency, FG, where fp is the fraction of the genome covered by the composite probe. We demonstrate the validity of this equation by showing that: (a) translocation detection efficiency predicted by the equation is consistent with experimental data as fp is changed; (b) translocation frequency dose-response curves measured in vitro using FISH agree well with dicentric frequency dose-response curves measured in vitro using conventional cytogenetic procedures; and (c) the genomic translocation frequencies estimated from FISH measurements for 20 Hiroshima A-bomb survivors and four workers exposed to ionizing radiation during the Y-12 criticality accident are approximately the same as the translocation frequencies measured using G-banding. We also show that translocation frequency dose response curves estimated using FISH are similar for Hiroshima A-bomb survivors and for first division lymphocytes irradiated in vitro. We conclude with a discussion of the potential utility of translocation frequency analysis for assessment of the level of acute radiation exposure independent of the time between analysis and exposure.